山萘酚抑制增生性瘢痕成纤维细胞的生长

Inhibition of hypertrophic scar fibroblasts growth by kaempferol

目的探讨山萘酚(kaempferol)对增生性瘢痕成纤维细胞生长的抑制作用。方法自2018年9月至2020年6月,北部战区总医院烧伤整形科从10例患者身上采集了10份增生性瘢痕组织样本。获取增生性瘢痕成纤维细胞后,分别使用0μmol/L(对照组)、50μmol/L、100μmol/L的山萘酚和10μmol/L的5-氟尿嘧啶(5-fluorouracil,5-FU)处理细胞,在处理后第0、1、2、3和4天使用CCK-8检测细胞的增殖情况。0μmol/L (对照组)、50μmol/L、100μmol/L的山萘酚和10μmol/L的5-FU作用于增生性瘢痕成纤维细胞1 d后,通过流式细胞技术检测不同细胞周期中的细胞占比,Western blot实验检测山萘酚对于Cyclin D1蛋白表达的作用;利用膜联蛋白V-碘化丙啶检测细胞的凋亡情况,Western blot检测bcl-2的表达情况。结果与对照组相比,不同时间点50、100μmol/L的山萘酚和10μmol/L的5-FU均能显著抑制增生性瘢痕成纤维细胞的增殖;山萘酚作用于增生性瘢痕成纤维细胞后流式细胞技术显示,其能够显著抑制细胞周期G1-S期的转化;Western blot结果显示,山萘酚和5-FU均能下调Cyclin D1的表达;膜联蛋白V-碘化丙啶结果显示,山萘酚和5-FU可以促进细胞凋亡比例;Western blot结果显示,山萘酚和5-FU能降低bcl-2的表达。结论山萘酚能够通过抑制Cyclink D1和bcl-2的表达发挥抑制增生性瘢痕成纤维细胞生长的作用。

Objective To explore the inhibitory effect of kaempferol on growth of hypertrophic scar fibroblasts. Methods From September 2018 to June 2020, 10 samples of hypertrophic scar were collected from 10 patients. The fibroblasts were treated by kaempferol of 0 μmol/L(control group), 50 μmol/L, 100 μmol/L and 5-fluorouracil of 10 μmol/L, respectively. At 0, 1, 2, 3 and 4 days after treatment,the proliferation of hypertrophic scar fibroblasts was detected by CCK-8 assay. After hypertrophic scar fibroblasts were treated by kaempferol of 0 μmol/L(control group), 50 μmol/L, 100 μmol/L and 5-fluorouracil of 10 μmol/L for 1 day, the cell proportion in different cell cycles was detected by flow cytometry. The effect of kaempferol on expression of Cyclin D1 protein and bcl-2 was detected by Western blot. The apoptosis of fibroblasts was detected by Annexin V-propidium iodide. Results Compared with the control group, the proliferation of hypertrophic scar fibroblasts at different time points could be significantly inhibited by kaempferol of 50 μmol/L, 100 μmol/L and5-fluorouracil of 10 μmol/L. After hypertrophic scar fibroblasts were treated by kaempferol, the flow cytometry demonstrated that the G1/S phase transition was significantly inhibited. The results of Western blot showed that the expression of cyclin D1 could be down regulated by kaempferol and 5-fluorouracil. The Annexin V-propidium iodide results showed that kaempferol and 5-fluorouracil could promote the apoptosis ratio of cells. Western blot results showed that kaempferol and 5-fluorouracil could reduce the expression of bcl-2. Conclusion The growth of hypertrophic scar fibroblasts could be inhibited by kaempferol.